GSM8046912: KO Th17 biol rep 2; Mus musculus; RNA-Seq
Total RNA was extracted for each of the 3 bioological replicates from WT Th17, KO Th17, WT Th17 with NAC, KO Th17 with NAC, WT Th17 with 3-KDS, KO Th17 with 3-KDS, WT naive and KO naive CD4+ T cells using Trizol and RNA-Clean and concentrater column. About 1.3 ug of RNA in 25ul was submitted for bulk RNAseq using Illumina machine (HWI-ST1276). Total RNA sample quality was measured using Nanodrop (OD260/280), gel electrophoresis and Agilent 2100 to measure RNA integrity. Susequently, mRNA was enriched using oligo(dT) beads, fragmented, cDNA was synthesized using random hexamers, followed by the second strand cDNA synthesis using dTTP. The library was checked with Qubit and real-time PCR for quantification and bioanalyzer for size distribution detection. Quantified libraries was pooled and sequenced on Illumina platforms, according to effective library concentration and data amount.